US2015037872A1PendingUtilityA1
Protein Variants Having Modified Immunogenicty
Est. expiryApr 28, 2020(expired)· nominal 20-yr term from priority
A61K 39/00C12Y 304/21062C12N 9/54A21D 2/267A21D 8/042C07K 1/047C07K 5/1008C07K 5/101C07K 5/1013C07K 5/1019C07K 5/1021C07K 14/001C07K 16/4283C11D 3/38627C11D 3/38645C12N 9/0061C12N 9/20C12Y 110/03002C07K 16/40Y10S435/911
72
PatentIndex Score
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Claims
Abstract
The present invention relates to a method of selecting a protein variant having modified immunogenicity as compared to the parent protein comprising the steps obtaining antibody binding peptide sequences, using the sequences to localise epitope sequences on the 3-dimensional structure of parent protein, defining an epitope area including amino acids situated within 5 Å from the epitope amino acids constituting the epitope sequence, changing one or more of the amino acids defining the epitope area of the parent protein by genetical engineering mutations of a DNA sequence encoding the parent protein, introducing the mutated DNA sequence into a suitable host, culturing said host and expressing the protein variant, and evaluating the immunogenicity of the protein variant using the parent protein as reference. The invention further relates to the protein variant and use thereof, as well as to a method for producing said protein variant.
Claims
exact text as granted — not AI-modified1 - 134 . (canceled)
135 . A protease variant having modified immunogenicity as compared to a parent protease, obtainable by a method comprising the steps of:
(a) obtaining antibody binding peptide sequences, (b) using the sequences to localise epitope sequences on the 3-dimensional structure of the parent protein, (c) defining an epitope area including amino acids situated within 5 Å from the epitope amino acids constituting the epitope sequence, (d) changing one or more of the amino acids defining the epitope area of the parent protein by genetic engineering mutations of a DNA sequence encoding the parent protein, (e) introducing the mutated DNA sequence into a suitable host, culturing said host and expressing the protein variant, and (f) evaluating the immunogenicity of the protein variant using the parent protein as reference.
136 . The protease variant of claim 135 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position −6 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position −5 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position −4 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position −2 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 3a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 28a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 44a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 44b to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 139 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 148 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 149 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion;
Position 264a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion.
137 . The protease variant of claim 135 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position −1 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, H;
Position 1 to V, L, I, W, M, F, Y, S, T, R;
Position 2 to G, V, I, M, F, N, Q, Y, S, T, H;
Position 3 to W, M, F, N, Q, Y, S, D, E, R, H;
Position 4 to V, L, W, M, F, Y, R;
Position 5 to V, L, I, W, M, F, N, Q, Y, T, R, H;
Position 6 to G, V, L, I, W, P, M, N, Q, T, D, E, R, H;
Position 9 to G, V, L, I, W, P, M, F, Q, Y, S, T, R, H;
Position 10 to G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R;
Position 12 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E;
Position 14 to V, L, I, W, P, M, F, N, Q, Y, T, R, H;
Position 15 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H;
Position 17 to G, A, V, I, W, P, M, F, Y, H;
Position 18 to G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H;
Position 19 to A, V, I, W, M, F, N, Y, S, T, D, R, H;
Position 20 to G, V, L, I, W, M, F, N, Q, Y, S, T, D, E;
Position 21 to G, V, I, W, N, Q, Y, S, T, D, E, R, H;
Position 22 to G, V, L, I, W, M, F, Y, S, T;
Position 24 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R;
Position 25 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position 27 to G, L, I, W, P, M, F, Y, T, H;
Position 38 to V, L, I, W, M, F, N, Q, Y, T, H;
Position 39 to G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H;
Position 40 to V, L, I, W, M, F, N, Q, Y, T, R, H;
Position 42 to G, A, L, W, C, M, F, N, Q, Y, S, T, D, E, R, H;
Position 43 to G, L, H;
Position 44 to G, V, L, I, W, P, M, F, Y, S, T;
Position 45 to G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 46 to G, A, L, I, W, P, M, F, Y, H;
Position 47 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 48 to A, L, I, P, M, F, N, Y, D, H;
Position 49 to G, A, V, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 50 to G, A, W, M, N, Q, Y, S, T, D, E, H;
Position 51 to V, L, I, W, M, F, N, Y, R;
Position 52 to V, L, I, W, M, F, Y, S, T, R;
Position 53 to A, V, L, I, W, M, F, N, Q, Y, S, D, E, H;
Position 54 to V, L, I, W, M, F, S, R;
Position 55 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R, K, H;
Position 56 to G, V, L, I, W, M, F, N, Q, Y, S, T, H;
Position 57 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 58 to L, W, M, F, N, Y, R;
Position 59 to A, V, L, I, C, T, H;
Position 61 to V, L, I, W, M, F, Y;
Position 62 to G, A, L, W, M, F, N, Y, R;
Position 64 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 75 to L;
Position 79 to I;
Position 80 to G;
Position 87 to A, V, L, I, W, M, F, Q, Y, S, T, D, E, H;
Position 89 to G, V, L, I, W, P, F, N, Y, T, E;
Position 91 to G, A, V, L, I, W, P, M, N, Y, S, T, D, E, R, H;
Position 98 to A;
Position 99 to V, L, I, W, M, F, Q, Y, H;
Position 100 to G, V, L, I, W, M, F, Y, R, H;
Position 101 to V, I, W, M, F, N, Q, Y, H;
Position 102 to V, L, I, W, M, F, Y, R, H, G;
Position 108 to I;
Position 109 to N;
Position 112 to E;
Position 113 to W;
Position 115 to I;
Position 117 to N;
Position 118 to N;
Position 126 to L;
Position 127 to G, A, V, I, W, M, F, Y, R, H, L;
Position 128 to I, W;
Position 129 to W;
Position 130 to W, F, Y, R;
Position 131 to W, Y, R;
Position 132 to L, W, M, F, Y, S, H;
Position 133 to A, L, I, W, M, F, Y, R;
Position 134 to L, I, W, F, N, Q, Y, R, H;
Position 136 to G, A, W, P, N, Y, S, T, D, E, H;
Position 137 to G, A, V, I, W, P, M, N, Y, H;
Position 140 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, H;
Position 141 to G, V, L, I, W, P, M, F, Q, S, D, E, H;
Position 143 to V, L, I, P, M, F, N, Y, R;
Position 144 to L, W, P, M, F, N, Q, Y, S, D, E, R, H;
Position 145 to G, V, L, I, W, M, F, Q, Y, D, E, R, H;
Position 146 to G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H;
Position 155 to V, L, I, W, M, F, Y, R;
Position 156 to V, I, W, F, R;
Position 157 to G, A, V, L, I, W, M, F, Y, T, R, H;
Position 158 to V, L, I, W, M, F, Y;
Position 159 to A, W, M, Y, T, R, H;
Position 160 to W, M, F, Y, R, H;
Position 161 to I, W, M, F, Y, H;
Position 167 to R, K;
Position 171 to D;
Position 172 to G, A, V, L, I, S, T, H;
Position 173 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H;
Position 181 to G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H;
Position 182 to A, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, H;
Position 183 to G, A, V, L, W, C, M, F, N, Q, Y, S, T, E, R, H;
Position 184 to A, V, L, I, W, C, M, F, N, Q, Y, T, E, H;
Position 185 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H;
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position 188 to G, A, V, L, W, F, S, R, K;
Position 189 to W, F;
Position 191 to A, V, L, I, W, M, F, Y, T, R, H;
Position 192 to G, L, I, W, M, N, Q, Y, S, T, D, R, H;
Position 194 to W, N, Q, Y, D, H;
Position 195 to W, P, Y;
Position 196 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 203 to V, F, Y, R, H;
Position 204 to I, W, M, Y, H;
Position 206 to F;
Position 209 to Y, R;
Position 210 to W, F, Y;
Position 211 to L, W, M, F, Y, H;
Position 212 to V, L, I, W, M, F, Y, T, R, H;
Position 214 to W, Y, R;
Position 215 to A, L, I, W, M, F, Y;
Position 216 to A, L, I, W, M, F, Y, R;
Position 217 to W, R;
Position 218 to G, A, L, W, P, M, F, Y, R, H;
Position 221 to S;
Position 236 to S;
Position 240 to N;
Position 241 to W;
Position 243 to N;
Position 245 to Q;
Position 247 to G, V, I, W, P, F, Y, S, T, R;
Position 248 to W, P, F, Y, E, R, H;
Position 249 to L, W, P, F, S, D, E, H;
Position 251 to G, L, I, W, P, M, F, Y, H;
Position 252 to G, A, W, P, N, Q, Y, T, E, R, H;
Position 254 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, H;
Position 255 to G, L, W, M, F, N, Y, T, D, H;
Position 256 to G, A, V, L, I, W, M, F, Q, Y, S, T, D, H;
Position 257 to G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H;
Position 258 to G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, E, K, H;
Position 259 to A, V, I, W, M, F, N, Q, Y, S, T, E, R;
Position 260 to L, I, W, M, F, Y, T, H;
Position 261 to L, N, S, H;
Position 262 to G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H;
Position 263 to G, A, V, L, I, P, C, M, N, Q, Y, S, T, R, K;
Position 265 to V, L, I, W, M, F, Y;
Position 269 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, R, H;
Position 271 to A, L, I, W, P, M, F, N, Y, S, T, R, H;
Position 272 to G, A, V, L, I, W, P, M, F, N, Q, Y, T, D, E, H;
Position 275 to G, A, V, L, I, W, M, F, N, Y, T, D.
138 . The protease variant of claim 135 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position −1 to Deletion;
Position 9 to Insertion, deletion;
Position 10 to Insertion, deletion;
Position 12 to Insertion, deletion;
Position 14 to Insertion, deletion;
Position 15 to Insertion, deletion;
Position 17 to Insertion, deletion;
Position 18 to Insertion, deletion;
Position 19 to Insertion, deletion;
Position 20 to Insertion, deletion;
Position 21 to Insertion, deletion;
Position 22 to Insertion, deletion;
Position 24 to Insertion, deletion;
Position 25 to Insertion, deletion;
Position 46 to Insertion, deletion;
Position 47 to Insertion, deletion;
Position 48 to Insertion, deletion;
Position 49 to Insertion, deletion;
Position 50 to Insertion, deletion;
Position 51 to Insertion, deletion;
Position 52 to Insertion, deletion;
Position 53 to Insertion, deletion;
Position 54 to Insertion, deletion;
Position 55 to Insertion, deletion;
Position 58 to Insertion, deletion;
Position 59 to Insertion, deletion;
Position 61 to Insertion, deletion;
Position 64 to Insertion, deletion;
Position 78 to Insertion:
Position 80 to Insertion;
Position 91 to Insertion, deletion;
Position 98 to Deletion;
Position 99 to Deletion;
Position 102 to Deletion;
Position 105 to Insertion;
Position 108 to Insertion;
Position 109 to Insertion;
Position 112 to Insertion;
Position 113 to Insertion;
Position 115 to Insertion;
Position 116 to Insertion;
Position 117 to Insertion;
Position 118 to Insertion;
Position 131 to Deletion;
Position 134 to Insertion, deletion;
Position 136 to Insertion, deletion;
Position 137 to Insertion, deletion;
Position 140 to Insertion, deletion;
Position 141 to Insertion, deletion;
Position 143 to Insertion, deletion;
Position 144 to Insertion, deletion;
Position 145 to Insertion, deletion;
Position 146 to Insertion, deletion;
Position 171 to Deletion;
Position 172 to Deletion;
Position 173 to Deletion;
Position 181 to Deletion;
Position 182 to Deletion;
Position 183 to Deletion;
Position 184 to Deletion;
Position 185 to Deletion;
Position 186 to Deletion;
Position 188 to Deletion;
Position 189 to Deletion;
Position 191 to Deletion;
Position 192 to Deletion;
Position 195 to Deletion;
Position 196 to Insertion, deletion;
Position 221 to Insertion;
Position 236 to Insertion;
Position 237 to Insertion;
Position 238 to Insertion;
Position 239 to Insertion;
Position 240 to Insertion;
Position 241 to Insertion;
Position 242 to Insertion;
Position 243 to Insertion;
Position 244 to Insertion;
Position 245 to Insertion;
Position 247 to Insertion, deletion;
Position 248 to Insertion, deletion;
Position 249 to Insertion, deletion;
Position 251 to Insertion, deletion;
Position 252 to Insertion, deletion;
Position 254 to Insertion, deletion;
Position 255 to Insertion, deletion;
Position 256 to Insertion, deletion;
Position 257 to Insertion, deletion;
Position 258 to Insertion, deletion;
Position 259 to Insertion, deletion;
Position 260 to Insertion, deletion;
Position 261 to Insertion, deletion;
Position 262 to Insertion, deletion;
Position 263 to Insertion, deletion;
Position 265 to Insertion, deletion;
Position 269 to Insertion, deletion;
Position 271 to Insertion, deletion;
Position 272 to Insertion, deletion;
Position 275 to Insertion, deletion.
139 . The protease variant of claim 135 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position 7 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 8 to G, A, L, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 13 to G, L, I, W, P, M, F, N, Q, Y, S, D, E, H;
Position 16 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, D, E, R, H;
Position 23 to G, A, V, L, I, W, M, F, Y, E, R, H;
Position 26 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position 28 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 29 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 33 to V, L, I, W, C, M, F, N, Q, Y, R, H;
Position 35 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 36 to V, L, I, W, P, M, F, N, Y, S, T, R, H;
Position 37 to L, I, W, M, F, N, Q, Y, S, R, H;
Position 41 to G, V, L, I, W, M, F, N, Q, Y, S, T, R, H;
Position 60 to G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H;
Position 63 to G, A, V, L, I, W, M, F, Y, T, R, H;
Position 73 to A;
Position 74 to A;
Position 81 to V;
Position 82 to L;
Position 86 to G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H;
Position 88 to A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position 92 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 93 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 94 to G, V, L, I, W, P, M, F, N, Y, T, D, E, K, H;
Position 96 to L, W, F, Y, R, K;
Position 97 to V, L, W, C, M, F, Y, H;
Position 111 to I;
Position 114 to A;
Position 119 to M;
Position 124 to M;
Position 135 to G, L, P, C, N, Q, T, R, H;
Position 138 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 142 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 147 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 151 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 162 to I, W, F, Y, R;
Position 163 to V, W, M, F, H;
Position 168 to G, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 169 to C, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y;
Position 174 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 176 to G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 179 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 187 to A, V, L, I, W, M, F, Y, R;
Position 190 to G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, R, K, H;
Position 193 to G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position 197 to G, V, L, I, W, P, M, F, Q, Y, S, T, H;
Position 198 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 205 to W, F, Y, R, K;
Position 208 to A, V, L, I, W, C, M, F, Y, T, R, K, H;
Position 219 to G, A, V, L, I, W, F, Y, R, H;
Position 222 to M;
Position 232 to A;
Position 233 to L;
Position 234 to I;
Position 250 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H;
Position 267 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H;
Position 268 to G, V, L, I, W, C, M, N, Q, Y, S, T, D, E, R, K, H;
Position 270 to G, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 273 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H;
Position 274 to W, P, M, F, N, Q, Y, T, D, E, R, H.
140 . The protease variant of claim 135 , wherein the protease is a subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position 13 to Insertion, deletion;
Position 16 to Insertion, deletion;
Position 23 to Insertion, deletion;
Position 26 to Insertion, deletion;
Position 28 to Insertion, deletion;
Position 29 to Insertion, deletion;
Position 35 to Deletion;
Position 60 to Insertion, deletion;
Position 63 to Insertion;
Position 81 to Insertion;
Position 82 to Insertion;
Position 92 to Insertion, deletion;
Position 93 to Insertion, deletion;
Position 94 to Insertion, deletion;
Position 96 to Deletion,
Position 106 to Insertion,
Position 111 to Insertion,
Position 114 to Insertion,
Position 119 to Insertion,
Position 124 to Insertion,
Position 138 to Insertion, deletion;
Position 142 to Insertion, deletion;
Position 147 to Insertion, deletion;
Position 151 to Insertion, deletion;
Position 174 to Insertion, deletion;
Position 176 to Insertion, deletion;
Position 179 to Insertion, deletion;
Position 187 to Deletion;
Position 190 to Deletion;
Position 193 to Deletion;
Position 197 to Insertion, deletion;
Position 198 to Insertion, deletion;
Position 232 to Insertion,
Position 233 to Insertion,
Position 234 to Insertion,
Position 246 to Insertion,
Position 250 to Insertion, deletion;
Position 267 to Insertion, deletion;
Position 268 to Insertion, deletion;
Position 270 to Insertion, deletion;
Position 273 to Insertion, deletion.
141 . The protease variant of claim 135 , wherein the protease is a savinase-like subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position 2 to G, V, I, M, F, N, Q, Y, S, T, H,
Position 3 to W, M, F, N, Q, Y, S, D, E, R, H,
Position 4 to V, L, W, M, F, Y, R,
Position 6 to G, V, L, I, W, P, M, N, Q, T, D, E, R, H,
Position 9 to G, V, L, I, W, P, M, F, Q, Y, S, T, R, H, insertion, deletion,
Position 10 to G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R, insertion, deletion,
Position 12 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position 14 to V, L, I, W, P, M, F, N, Q, Y, T, R, H, insertion, deletion,
Position 15 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H, insertion, deletion,
Position 17 to G, A, V, I, W, P, M, F, Y, H, insertion, deletion,
Position 18 to G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position 19 to A, V, I, W, M, F, N, Y, S, T, D, R, H, insertion, deletion,
Position 20 to G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position 21 to G, V, I, W, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 22 to G, V, L, I, W, M, F, Y, S, T, insertion, deletion,
Position 24 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, insertion, deletion,
Position 25 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 27 to G, L, I, W, P, M, F, Y, T, H,
Position 37 to L, I, W, M, F, N, Q, Y, S, R, H,
Position 40 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 42 to G, A, L, W, C, M, F, N, Q, Y, S, T, D, E, R, H,
Position 43 to G, L, H,
Position 44 to G, V, L, I, W, P, M, F, Y, S, T,
Position 45 to G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H,
Position 46 to G, A, L, I, W, P, M, F, Y, H, insertion, deletion,
Position 47 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 48 to A, L, I, P, M, F, N, Y, D, H, insertion, deletion,
Position 50 to G, A, W, M, N, Q, Y, S, T, D, E, H, insertion, deletion,
Position 51 to V, L, I, W, M, F, N, Y, R, deletion, insertion,
Position 54 to V, L, I, W, M, F, S, R, deletion, insertion,
Position 55 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R, K, H, deletion, insertion,
Position 57 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H,
Position 58 to L, W, M, F, N, Y, R, insertion, deletion,
Position 59 to A, V, L, I, C, T, H, insertion, deletion,
Position 61 to V, L, I, W, M, F, Y, insertion, deletion,
Position 64 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 75 to L,
Position 78 to insertion,
Position 79 to I,
Position 87 to A, V, L, I, W, M, F, Q, Y, S, T, D, E, H,
Position 89 to G, V, L, I, W, P, F, N, Y, T, E,
Position 91 to G, A, V, L, I, W, P, M, N, Y, S, T, D, E, R, H, insertion, deletion,
Position 98 to A, deletion,
Position 100 to G, V, L, I, W, M, F, Y, R, H,
Position 101 to V, I, W, M, F, N, Q, Y, H,
Position 102 to V, L, I, W, M, F, Y, R, H, G, deletion,
Position 109 to N, insertion,
Position 112 to E, insertion,
Position 113 to W, insertion,
Position 116 to insertion,
Position 117 to N, insertion,
Position 126 to L,
Position 127 to G, A, V, I, W, M, F, Y, R, H, L,
Position 128 to I, W,
Position 129 to W,
Position 130 to W, F, Y, R,
Position 131 to W, Y, R, deletion,
Position 132 to L, W, M, F, Y, S, H,
Position 133 to A, L, I, W, M, F, Y, R,
Position 134 to L, I, W, F, N, Q, Y, R, H, insertion, deletion,
Position 136 to G, A, W, P, N, Y, S, T, D, E, H, insertion, deletion,
Position 137 to G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position 140 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, H, insertion, deletion,
Position 141 to G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position 143 to V, L, I, P, M, F, N, Y, R, insertion, deletion,
Position 144 to L, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 145 to G, V, L, I, W, M, F, Q, Y, D, E, R, H, insertion, deletion,
Position 146 to G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 155 to V, L, I, W, M, F, Y, R,
Position 156 to V, I, W, F, R,
Position 157 to G, A, V, L, I, W, M, F, Y, T, R, H,
Position 158 to V, L, I, W, M, F, Y,
Position 160 to W, M, F, Y, R, H,
Position 161 to I, W, M, F, Y, H,
Position 167 to R, K,
Position 170 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position 171 to D, deletion,
Position 172 to G, A, V, L, I, S, T, H, deletion,
Position 173 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H, deletion,
Position 181 to G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H, deletion,
Position 183 to G, A, V, L, W, C, M, F, N, Q, Y, S, T, E, R, H, deletion,
Position 184 to A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position 185 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 188 to G, A, V, L, W, F, S, R, K, deletion,
Position 189 to W, F, deletion,
Position 191 to A, V, L, I, W, M, F, Y, T, R, H, deletion,
Position 192 to G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position 194 to W, N, Q, Y, D, H,
Position 195 to W, P, Y, deletion,
Position 197 to G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position 203 to V, F, Y, R, H,
Position 206 to F,
Position 209 to Y, R,
Position 210 to W, F, Y,
Position 212 to V, L, I, W, M, F, Y, T, R, H,
Position 214 to W, Y, R,
Position 216 to A, L, I, W, M, F, Y, R,
Position 217 to W, R,
Position 218 to G, A, L, W, P, M, F, Y, R, H,
Position 221 to S, insertion,
Position 236 to S, insertion,
Position 237 to insertion,
Position 239 to insertion,
Position 240 to N, insertion,
Position 241 to W, insertion,
Position 242 to insertion,
Position 244 to insertion,
Position 245 to Q, insertion,
Position 247 to G, V, I, W, P, F, Y, S, T, R, insertion, deletion,
Position 248 to W, P, F, Y, E, R, H, insertion, deletion,
Position 251 to G, L, I, W, P, M, F, Y, H, insertion, deletion,
Position 252 to G, A, W, P, N, Q, Y, T, E, R, H, insertion, deletion,
Position 255 to G, L, W, M, F, N, Y, T, D, H, insertion, deletion,
Position 256 to G, A, V, L, I, W, M, F, Q, Y, S, T, D, H, insertion, deletion,
Position 257 to G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position 258 to G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, E, K, H, insertion, deletion,
Position 259 to A, V, I, W, M, F, N, Q, Y, S, T, E, R, insertion, deletion,
Position 260 to L, I, W, M, F, Y, T, H, insertion, deletion,
Position 261 to L, N, S, H, insertion, deletion,
Position 262 to G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 263 to G, A, V, L, I, P, C, M, N, Q, Y, S, T, R, K, insertion, deletion,
Position 265 to V, L, I, W, M, F, Y, insertion, deletion,
Position 271 to A, L, I, W, P, M, F, N, Y, S, T, R, H, insertion, deletion,
Position 272 to G, A, V, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position 275 to G, A, V, L, I, W, M, F, N, Y, T, D, insertion, deletion.
142 . The protease variant of claim 141 , wherein the savinase-like subtilisin comprises one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position 6 to G, V, L, I, W, P, M, N, Q, T, D, E, R, H,
Position 9 to G, V, L, I, W, P, M, F, Q, Y, S, T, R, H, insertion, deletion,
Position 10 to G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R, insertion, deletion,
Position 14 to V, L, I, W, P, M, F, N, Q, Y, T, R, H, insertion, deletion,
Position 15 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H, insertion, deletion,
Position 17 to G, A, V, I, W, P, M, F, Y, H, insertion, deletion,
Position 18 to G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position 19 to A, V, I, W, M, F, N, Y, S, T, D, R, H, insertion, deletion,
Position 20 to G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position 21 to G, V, I, W, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 37 to L, I, W, M, F, N, Q, Y, S, R, H,
Position 43 to G, L, H,
Position 45 to G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H,
Position 47 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 50 to G, A, W, M, N, Q, Y, S, T, D, E, H, insertion, deletion,
Position 51 to V, L, I, W, M, F, N, Y, R, deletion, insertion,
Position 54 to V, L, I, W, M, F, S, R, deletion, insertion,
Position 59 to A, V, L, I, C, T, H, insertion, deletion,
Position 89 to G, V, L, I, W, P, F, N, Y, T, E,
Position 91 to G, A, V, L, I, W, P, M, N, Y, S, T, D, E, R, H, insertion, deletion,
Position 101 to V, I, W, M, F, N, Q, Y, H,
Position 109 to N, insertion,
Position 112 to E, insertion,
Position 113 to W, insertion,
Position 127 to G, A, V, I, W, M, F, Y, R, H, L,
Position 128 to I, W,
Position 129 to W,
Position 130 to W, F, Y, R,
Position 131 to W, Y, R, deletion,
Position 133 to A, L, I, W, M, F, Y, R,
Position 136 to G, A, W, P, N, Y, S, T, D, E, H, insertion, deletion,
Position 137 to G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position 140 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, H, insertion, deletion,
Position 141 to G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position 143 to V, L, I, P, M, F, N, Y, R, insertion, deletion,
Position 144 to L, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 145 to G, V, L, I, W, M, F, Q, Y, D, E, R, H, insertion, deletion,
Position 146 to G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 155 to V, L, I, W, M, F, Y, R,
Position 157 to G, A, V, L, I, W, M, F, Y, T, R, H,
Position 158 to V, L, I, W, M, F, Y,
Position 160 to W, M, F, Y, R, H,
Position 161 to I, W, M, F, Y, H,
Position 167 to R, K,
Position 170 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position 171 to D, deletion,
Position 172 to G, A, V, L, I, S, T, H, deletion,
Position 173 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H, deletion,
Position 181 to G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H, deletion,
Position 184 to A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position 185 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 188 to G, A, V, L, W, F, S, R, K, deletion,
Position 189 to W, F, deletion,
Position 192 to G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position 194 to W, N, Q, Y, D, H,
Position 195 to W, P, Y, deletion,
Position 197 to G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position 203 to V, F, Y, R, H,
Position 210 to W, F, Y,
Position 218 to G, A, L, W, P, M, F, Y, R, H,
Position 236 to S, insertion,
Position 237 to insertion,
Position 239 to insertion,
Position 240 to N, insertion,
Position 241 to W, insertion,
Position 242 to insertion,
Position 244 to insertion,
Position 245 to Q, insertion,
Position 247 to G, V, I, W, P, F, Y, S, T, R, insertion, deletion,
Position 251 to G, L, I, W, P, M, F, Y, H, insertion, deletion,
Position 255 to G, L, W, M, F, N, Y, T, D, H, insertion, deletion,
Position 256 to G, A, V, L, I, W, M, F, Q, Y, S, T, D, H, insertion, deletion,
Position 257 to G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position 258 to G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, E, K, H, insertion, deletion,
Position 260 to L, I, W, M, F, Y, T, H, insertion, deletion,
Position 262 to G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 265 to V, L, I, W, M, F, Y, insertion, deletion,
Position 271 to A, L, I, W, P, M, F, N, Y, S, T, R, H, insertion, deletion,
Position 272 to G, A, V, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position 275 to G, A, V, L, I, W, M, F, N, Y, T, D, insertion, deletion.
143 . The savinase-like subtilisin of claim 141 , wherein the subtilisin has any of the amino acid sequence of SEQ ID NO: 24, 26, 27, 28, 29, 30, 31, 32, 34, 35.
144 . The protein variant of claim 135 , wherein the protease is a savinase-like subtilisin comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position 8 to G, A, L, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H,
Position 16 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 23 to G, A, V, L, I, W, M, F, Y, E, R, H, insertion, deletion,
Position 26 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 35 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H, deletion,
Position 38 to V, L, I, W, M, F, N, Q, Y, T, H,
Position 39 to G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H,
Position 41 to G, V, L, I, W, M, F, N, Q, Y, S, T, R, H,
Position 60 to G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H, insertion, deletion,
Position 73 to A,
Position 74 to A,
Position 80 to G, insertion,
Position 81 to V, insertion,
Position 86 to G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H,
Position 88 to A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H,
Position 90 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 93 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 108 to I, insertion,
Position 111 to I, insertion,
Position 124 to M, insertion,
Position 135 to G, L, P, C, N, Q, T, R, H,
Position 142 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 147 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 148 to G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 149 to G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 151 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 163 to V, W, M, F, H,
Position 168 to G, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, R, K, H,
Position 169 to C, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y,
Position 174 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 179 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 190 to G, A, V, L, I, W, C, M, F, N, Q, Y, S, T, R, K, H, deletion,
Position 193 to G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 196 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 208 to A, V, L, I, W, C, M, F, Y, T, R, K, H,
Position 213 to N, oN, E,
Position 215 to A, L, I, W, M, F, Y,
Position 232 to A, insertion,
Position 233 to L, insertion,
Position 234 to I, insertion,
Position 246 to insertion,
Position 250 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 254 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 267 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 268 to G, V, L, I, W, C, M, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 269 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, R, H, insertion, deletion,
Position 273 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion.
145 . The savinase-like subtilisin of claim 144 , wherein the subtilisin has any of the amino acid sequence of SEQ ID NO: 24, 26, 27, 28, 29, 30, 31, 32, 34, 35.
146 . The protease variant of claim 135 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 25 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 25:
Position 21 to G, V, I, W, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 27 to G, L, I, W, P, M, F, Y, T, H,
Position 50 to G, A, W, M, N, Q, Y, S, T, D, E, H, insertion, deletion,
Position 52 to V, L, I, W, M, F, Y, S, T, R, deletion, insertion,
Position 55 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R, K, H, deletion, insertion,
Position 129 to W,
Position 133 to A, L, I, W, M, F, Y, R,
Position 172 to G, A, V, L, I, S, T, H, deletion,
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 194 to W, N, Q, Y, D, H,
Position 195 to W, P, Y, deletion,
Position 197 to G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position 242 to insertion,
Position 249 to L, W, P, F, S, D, E, H, insertion, deletion,
Position 252 to G, A, W, P, N, Q, Y, T, E, R, H, insertion, deletion,
Position 254 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 257 to G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position 260 to L, I, W, M, F, Y, T, H, insertion, deletion,
Position 265 to V, L, I, W, M, F, Y, insertion, deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
147 . The protein variant of claim 135 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 10 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 10:
Position 4 to V, L, W, M, F, Y, R,
Position 38 to V, L, I, W, M, F, N, Q, Y, T, H,
Position 40 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 43 to G, L, H,
Position 47 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 49 to G, A, V, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 54 to V, L, I, W, M, F, S, R, deletion, insertion,
Position 96 to L, W, F, Y, R, K, deletion,
Position 99 to V, L, I, W, M, F, Q, Y, H, deletion,
Position 113 to W, insertion,
Position 131 to W, Y, R, deletion,
Position 133 to A, L, I, W, M, F, Y, R,
Position 137 to G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position 141 to G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position 144 to L, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 170 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y;
Position 173 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, E, H, deletion,
Position 181 to G, A, V, L, I, W, C, M, F, Q, Y, T, D, R, K, H, deletion,
Position 185 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, E, H, deletion,
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 188 to G, A, V, L, W, F, S, R, K, deletion,
Position 194 to W, N, Q, Y, D, H,
Position 203 to V, F, Y, R, H,
Position 210 to W, F, Y,
Position 211 to L, W, M, F, Y, H,
Position 257 to G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position 261 to L, N, S, H, insertion, deletion,
Position 262 to G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 265 to V, L, I, W, M, F, Y, insertion, deletion.
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
148 . The protein variant of claim 135 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 11 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 11:
Position 38 to V, L, I, W, M, F, N, Q, Y, T, H,
Position 40 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 45 to G, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H,
Position 47 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 49 to G, A, V, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 50 to G, A, W, M, N, Q, Y, S, T, D, E, H, insertion, deletion,
Position 52 to V, L, I, W, M, F, Y, S, T, R, deletion, insertion,
Position 53 to A, V, L, I, W, M, F, N, Q, Y, S, D, E, H, deletion, insertion,
Position 56 to G, V, L, I, W, M, F, N, Q, Y, S, T, H,
Position 58 to L, W, M, F, N, Y, R, insertion, deletion,
Position 96 to L, W, F, Y, R, K, deletion,
Position 97 to V, L, W, C, M, F, Y, H,
Position 98 to A, deletion,
Position 105 to insertion,
Position 109 to N, insertion,
Position 113 to W, insertion,
Position 115 to I, insertion,
Position 133 to A, L, I, W, M, F, Y, R,
Position 136 to G, A, W, P, N, Y, S, T, D, E, H, insertion, deletion,
Position 137 to G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position 141 to G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position 158 to V, L, I, W, M, F, Y,
Position 159 to A, W, M, Y, T, R, H,
Position 172 to G, A, V, L, I, S, T, H, deletion,
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 189 to W, F, deletion,
Position 192 to G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position 195 to W, P, Y, deletion,
Position 197 to G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position 257 to G, A, L, I, W, C, M, F, N, Q, Y, S, T, D, E, K, H, insertion, deletion,
Position 261 to L, N, S, H, insertion, deletion,
Position 265 to V, L, I, W, M, F, Y, insertion, deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
149 . The protein variant of claim 135 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 33 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 33:
Position −6 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, Y, insertion, deletion,
Position −5 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, T, V, W, Y, insertion, deletion,
Position −4 to A, C, D, E, F, G, H, I, K, L, M, N, Q, R, S, T, V, W, Y, insertion, deletion,
Position −2 to A, C, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position −1 to G, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 1 to V, L, I, W, M, F, Y, S, T, R,
Position 2 to G, V, I, M, F, N, Q, Y, S, T, H,
Position 3a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 5 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 6 to G, V, L, I, W, P, M, N, Q, T, D, E, R, H,
Position 7 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H,
Position 8 to G, A, L, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H,
Position 10 to G, A, V, I, W, P, M, N, Q, Y, S, T, D, E, R, insertion, deletion,
Position 12 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, insertion, deletion,
Position 13 to G, L, I, W, P, M, F, N, Q, Y, S, D, E, H, insertion, deletion,
Position 14 to V, L, I, W, P, M, F, N, Q, Y, T, R, H, insertion, deletion,
Position 15 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, E, H, insertion, deletion,
Position 16 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 17 to G, A, V, I, W, P, M, F, Y, H, insertion, deletion,
Position 18 to G, A, L, I, W, P, M, F, N, Q, Y, T, D, E, H, insertion, deletion,
Position 19 to A, V, I, W, M, F, N, Y, S, T, D, R, H, insertion, deletion,
Position 21 to G, V, I, W, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 22 to G, V, L, I, W, M, F, Y, S, T, insertion, deletion,
Position 23 to G, A, V, L, I, W, M, F, Y, E, R, H, insertion, deletion,
Position 24 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, insertion, deletion,
Position 25 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 26 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 27 to G, L, I, W, P, M, F, Y, T, H,
Position 28 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 28a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 29 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 33 to V, L, I, W, C, M, F, N, Q, Y, R, H,
Position 35 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H, deletion,
Position 37 to L, I, W, M, F, N, Q, Y, S, R, H,
Position 40 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 42 to G, A, L, W, C, M, F, N, Q, Y, S, T, D, E, R, H,
Position 43 to G, L, H,
Position 44 to G, V, L, I, W, P, M, F, Y, S, T,
Position 44a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 44b to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 46 to G, A, L, I, W, P, M, F, Y, H, insertion, deletion,
Position 48 to A, L, I, P, M, F, N, Y, D, H, insertion, deletion,
Position 51 to V, L, I, W, M, F, N, Y, R, deletion, insertion,
Position 52 to V, L, I, W, M, F, Y, S, T, R, deletion, insertion,
Position 53 to A, V, L, I, W, M, F, N, Q, Y, S, D, E, H, deletion, insertion,
Position 55 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R, K, H, deletion, insertion,
Position 56 to G, V, L, I, W, M, F, N, Q, Y, S, T, H,
Position 57 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, K, H,
Position 58 to L, W, M, F, N, Y, R, insertion, deletion,
Position 61 to V, L, I, W, M, F, Y, insertion, deletion,
Position 64 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 75 to L,
Position 81 to insertion,
Position 86 to G, A, V, L, I, W, M, F, N, Q, Y, T, D, E, R, H,
Position 87 to A, V, L, I, W, M, F, Q, Y, S, T, D, E, H,
Position 88 to A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H,
Position 89 to G, V, L, I, W, P, F, N, Y, T, E,
Position 91 to G, A, V, L, I, W, P, M, N, Y, S, T, D, E, R, H, insertion, deletion,
Position 92 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 94 to G, V, L, I, W, P, M, F, N, Y, T, D, E, K, H, insertion, deletion,
Position 96 to L, W, F, Y, R, K, deletion,
Position 97 to V, L, W, C, M, F, Y, H,
Position 98 to deletion,
Position 101 to V, I, W, M, F, N, Q, Y, H,
Position 102 to V, L, I, W, M, F, Y, R, H, G, deletion,
Position 108 to I, insertion,
Position 109 to N, insertion,
Position 111 to insertion,
Position 112 to E, insertion,
Position 113 to W, insertion,
Position 114 to insertion,
Position 115 to I, insertion,
Position 117 to N, insertion,
Position 118 to N, insertion,
Position 119 to M, insertion,
Position 127 to G, A, V, I, W, M, F, Y, R, H, L,
Position 133 to A, L, I, W, M, F, Y, R,
Position 134 to L, I, W, F, N, Q, Y, R, H, insertion, deletion,
Position 135 to G, L, P, C, N, Q, T, R, H,
Position 136 to G, A, W, P, N, Y, S, T, D, E, H, insertion, deletion,
Position 137 to G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position 138 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 139 to G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 141 to G, V, L, I, W, P, M, F, Q, S, D, E, H, insertion, deletion,
Position 142 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 144 to L, W, P, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 145 to G, V, L, I, W, M, F, Q, Y, D, E, R, H, insertion, deletion,
Position 146 to G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 147 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 148 to G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 156 to V, I, W, F, R,
Position 158 to V, L, I, W, M, F, Y,
Position 160 to W, M, F, Y, R, H,
Position 161 to I, W, M, F, Y, H,
Position 162 to I, W, F, Y, R,
Position 163 to V, W, M, F, H,
Position 167 to R, K,
Position 169 to C, E, F, G, H, I, K, L, M, N, Q, R, T, V, W, Y,
Position 170 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 171 to D, deletion,
Position 174 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 176 to G, A, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 182 to A, V, L, I, W, C, M, F, N, Q, Y, S, T, D, E, H, deletion,
Position 186 to G, A, V, L, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 188 to G, A, V, L, W, F, S, R, K, deletion,
Position 191 to A, V, L, I, W, M, F, Y, T, R, H, deletion,
Position 192 to G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position 193 to G, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, deletion,
Position 194 to W, N, Q, Y, D, H,
Position 195 to W, P, Y, deletion,
Position 196 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 197 to G, V, L, I, W, P, M, F, Q, Y, S, T, H, insertion, deletion,
Position 198 to G, A, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 203 to V, F, Y, R, H,
Position 205 to W, F, Y, R, K,
Position 215 to A, L, I, W, M, F, Y,
Position 216 to A, L, I, W, M, F, Y, R,
Position 217 to W, R,
Position 219 to G, A, V, L, I, W, F, Y, R, H,
Position 233 to insertion,
Position 234 to I, insertion,
Position 236 to insertion,
Position 237 to insertion,
Position 238 to insertion,
Position 239 to insertion,
Position 240 to insertion,
Position 243 to insertion,
Position 246 to insertion,
Position 247 to G, V, I, W, P, F, Y, S, T, R, insertion, deletion,
Position 249 to L, W, P, F, S, D, E, H, insertion, deletion,
Position 252 to G, A, W, P, N, Q, Y, T, E, R, H, insertion, deletion,
Position 254 to G, V, L, I, W, M, F, N, Q, Y, S, D, E, R, H, insertion, deletion,
Position 262 to G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 264a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
Position 270 to G, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 273 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 274 to W, P, M, F, N, Q, Y, T, D, E, R, H,
Position 275 to G, A, V, L, I, W, M, F, N, Y, T, D, insertion, deletion,
Position 276 to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
150 . The protein variant of claim 135 having modified immunogenicity as compared to its parent protein having at least 81% homology to SEQ ID NO: 33 comprising one or more of the following substitutions corresponding to any of the following in SEQ ID NO: 33:
Position 5 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 22 to G, V, L, I, W, M, F, Y, S, T, insertion, deletion,
Position 26 to G, A, V, L, I, W, M, F, N, Q, Y, S, T, D, E, R, H, insertion, deletion,
Position 28 to G, A, V, L, I, W, P, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 37 to L, I, W, M, F, N, Q, Y, S, R, H,
Position 40 to V, L, I, W, M, F, N, Q, Y, T, R, H,
Position 44 to G, V, L, I, W, P, M, F, Y, S, T,
Position 51 to V, L, I, W, M, F, N, Y, R, deletion, insertion,
Position 52 to V, L, I, W, M, F, Y, S, T, R, deletion, insertion,
Position 55 to G, A, V, L, I, W, C, M, F, N, Q, Y, T, D, E, R, K, H, deletion, insertion,
Position 58 to L, W, M, F, N, Y, R, insertion, deletion,
Position 61 to V, L, I, W, M, F, Y, insertion, deletion,
Position 64 to G, V, L, I, W, P, C, M, F, N, Q, Y, S, T, D, E, R, K, H, insertion, deletion,
Position 87 to A, V, L, I, W, M, F, Q, Y, S, T, D, E, H,
Position 97 to V, L, W, C, M, F, Y, H,
Position 98 to deletion,
Position 101 to V, I, W, M, F, N, Q, Y, H,
Position 102 to V, L, I, W, M, F, Y, R, H, G, deletion,
Position 109 to N, insertion,
Position 112 to E, insertion,
Position 118 to N, insertion,
Position 127 to G, A, V, I, W, M, F, Y, R, H, L,
Position 137 to G, A, V, I, W, P, M, N, Y, H, insertion, deletion,
Position 146 to G, A, W, L, I, W, M, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 156 to V, I, W, F, R,
Position 158 to V, L, I, W, M, F, Y,
Position 161 to I, W, M, F, Y, H,
Position 188 to G, A, V, L, W, F, S, R, K, deletion,
Position 192 to G, L, I, W, M, N, Q, Y, S, T, D, R, H, deletion,
Position 194 to W, N, Q, Y, D, H,
Position 195 to W, P, Y, deletion,
Position 203 to V, F, Y, R, H,
Position 216 to A, L, I, W, M, F, Y, R,
Position 236 to insertion,
Position 237 to insertion,
Position 262 to G, A, V, L, I, W, P, F, N, Q, Y, T, D, E, R, H, insertion, deletion,
Position 264a to A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, Y, insertion, deletion,
with the proviso that the amino acids of the parent enzyme are substituted to another amino acid.
151 . A composition comprising the protein variant of claim 135 .
152 . A DNA construct comprising a DNA sequence encoding a protein variant of claim 135 .
153 . A host cell which is capable of expressing a polypeptide and comprising a DNA construct of claim 152 .
154 . A method of producing a protein variant having reduced immunogenicity as compared to the parent protein, comprising:
(a) culturing the host cell of claim 153 in a suitable culture medium to obtain expression and secretion of the protein into the medium, followed by (b) isolation of the protein from the culture medium.Cited by (0)
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